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北京大学学报（医学版） ›› 2024, Vol. 56 ›› Issue (5): 860-867. doi: 10.19723/j.issn.1671-167X.2024.05.017

• 论著 • 上一篇    下一篇

miR-125b-5p修饰脐带间充质干细胞对系统性红斑狼疮的免疫调控机制

武志慧1, 胡明智1, 赵巧英1, 吕凤凤1, 张晶莹2, 张伟1, 王永福2, 孙晓林1,*(), 王慧2,*()   

  1. 1. 包头医学院第一附属医院中心实验室(内蒙古自治区自体免疫学重点实验室)，内蒙古自治区包头 014010
    2. 包头医学院第一附属医院风湿免疫科，内蒙古自治区包头 014010
  • 收稿日期:2021-07-26 出版日期:2024-10-18 发布日期:2024-10-16
  • 通讯作者: 孙晓林,王慧 E-mail:yolo_1010@126.com;wanghuier@126.com
  • 基金资助:
    国家自然科学基金(81860294);国家自然科学基金(81860295);内蒙古自治区自然科学基金(2019MS08055);内蒙古自治区科技计划项目(201802089);内蒙古自治区科技计划项目(2019GG052)

Immunomodulatory mechanism of umbilical cord mesenchymal stem cells modified by miR-125b-5p in systemic lupus erythematosus

Zhihui WU1, Mingzhi HU1, Qiaoying ZHAO1, Fengfeng LV1, Jingying ZHANG2, Wei ZHANG1, Yongfu WANG2, Xiaolin SUN1,*(), Hui WANG2,*()   

  1. 1. Central Laboratory, First Affiliated Hospital of Baotou Medical College (Inner Mongolia Key Laboratory of Autoimmunology), Baotou 014010, Inner Mongolia Autonomous Region, China
    2. Department of Rheumatism and Immunology, First Affiliated Hospital of Baotou Medical College, Baotou 014010, Inner Mongolia Autonomous Region, China
  • Received:2021-07-26 Online:2024-10-18 Published:2024-10-16
  • Contact: Xiaolin SUN, Hui WANG E-mail:yolo_1010@126.com;wanghuier@126.com
  • Supported by:
    Supported by the National Natural Science Foundation of China(81860294);Supported by the National Natural Science Foundation of China(81860295);the Natural Science Foundation of Inner Mongolia(2019MS08055);the Inner Mongolia Autonomous Region Science and Technology Plan Projects(201802089);the Inner Mongolia Autonomous Region Science and Technology Plan Projects(2019GG052)

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摘要：

目的: 研究miR-125b-5p修饰脐带间充质干细胞(umbilical cord mesenchymal stem cells，UC-MSCs)对系统性红斑狼疮(systemic lupus erythematosus，SLE)的免疫调控作用机制。方法: 实时荧光定量PCR检测miR-125b-5p在UC-MSCs和SLE患者、健康体检者外周血单个核细胞中的表达水平；Annexin V-FITC/PI凋亡检测试剂盒检测miR-125b-5p对UC-MSCs的凋亡影响；每周对MRL/lpr小鼠进行尾静脉注射UC-MSCs，5周后流式细胞术检测各组小鼠脾细胞T淋巴细胞亚群分化情况；ELISA法检测各组MRL/lpr小鼠血清中白细胞介素(interleukin，IL)-4和IL-17A表达水平；苏木精-伊红染色法观察MRL/lpr小鼠肺和肾组织病理变化。结果: 与健康对照组相比，SLE患者外周血单个核细胞中miR-125b-5p的表达水平显著下调(P < 0.01)；与UC-MSCs相比，miR-125b-5p转染UC-MSCs组miR-125b-5p的表达水平显著上调(P < 0.01)。miR-125b-5p转染UC-MSCs 48 h可显著提高细胞存活率(P < 0.01)；与未处理的MRL/lpr小鼠组相比，miR-125b-5p修饰UC-MSCs对MRL/lpr小鼠脾脏Th17细胞分化具有明显的下调作用(P < 0.05)，且在小鼠血清中IL-4表达水平明显升高(P < 0.05)，IL-17A表达水平明显减低(P < 0.05)。经miR-125b-5p修饰的UC-MSCs治疗后，MRL/lpr小鼠肺和肾组织内炎性细胞浸润和微血栓减少。结论: miR-125b-5p修饰UC-MSCs对SLE具有免疫调控作用。

关键词: 间充质干细胞, 基因修饰, 系统性红斑狼疮, 免疫调节, 脐带

Abstract:

Objective: To investigate the mechanism of immunomodulatory effects of umbilical cord mesenchymal stem cells (UC-MSCs) modified by miR-125b-5p on systemic lupus erythematosus (SLE). Methods: The expression level of miR-125b-5p was detected by real-time fluorescence quantitative PCR in UC-MSCs and peripheral blood mononuclear cells (PBMCs) from SLE patients and health checkers. Annexin V-FITC/PI apoptosis detection kit was used to detect the effect of miR-125b-5p on apoptosis of UC-MSCs. MRL/lpr mice in each group were injected with UC-MSCs via tail vein, and T-lymphocyte subsets in the spleen of the MRL/lpr mice were detected by flow cytometry after 5 weeks. The expression levels of interleukin (IL)-4 and IL-17A in serum of MRL/lpr mice were detected by ELISA. Hematoxylin-eosin staining was used to observe the pathological manifestations of the lungs and kidneys of the MRL/lpr mice. Results: miR-125b-5p was significantly down-regulated in PBMCs of SLE patients compared with healthy controls (P < 0.01). Compared with the UC-MSCs group, the expression of miR- 125b-5p in UC-MSCs modified by miR-125b-5p group was increased (P < 0.01). The survival rate of UC-MSCs was significantly increased by miR-125b-5p (P < 0.01). Compared with the untreated group of MRL/lpr mice, the expression level of IL-4 in serum was increased (P < 0.05); the expression level of IL-17A was decreased (P < 0.05); the proportion of Th17 cells in the spleen of MRL/lpr mice was decreased (P < 0.05); the inflammatory cells infiltration and micro-thrombosis of lungs and kidneys of MRL/lpr mice were significantly reduced in the UC-MSCs modified by miR-125b-5p treatment group. Conclusion: UC-MSCs modified by miR-125b-5p have immunomodulatory effects on systemic lupus erythematosus.

Key words: Mesenchymal stem cells, Gene modification, Systemic lupus erythematosus, Immunomo-dulation, Umbilical cord

中图分类号: 

  • R593.2

图1

健康对照和SLE患者PBMCs中miR-125b-5p的表达水平(n=19)"

图2

miR-125b-5p在转染后的UC-MSCs中的表达(n=3)"

图3

各组UC-MSCs凋亡率"

图4

MRL/lpr小鼠脾细胞Th17、Treg、Th1、Th2细胞流式检测图"

图5

各组MRL/lpr小鼠脾细胞T细胞亚群分化情况(n=4)"

表1

各组MRL/lpr小鼠血清IL-4、IL-17A表达水平(n=4)"

Items Control PBS UC-MSCs UC-MSCs+miR-NC UC-MSCs+miR-125b-5p F P
IL-4/(ng/L) 242.53±5.38 247.13±8.40 250.55±5.67 249.22±4.14 251.94±6.02* 1.456 0.264
IL-17A/(ng/L) 131.65±20.04 113.85±26.17 143.03±14.90 139.14±5.38 101.23±12.90* 4.174 0.018

图6

各组MRL/lpr小鼠肺组织及肾组织病理形态(HE ×22.9)"

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ISSN 1671-167X
CN 11-4691/R
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